SLC15A1 Knockout HuH-7 Cell Line

The choice depends on whether you are studying SLC15A1 (PEPT1)'s role as the principal intestinal di/tripeptide transporter or its functions in oral drug bioavailability. The Knockout line is the standard tool for asking whether PEPT1 is required for cellular uptake of dipeptides, tripeptides, and peptide-mimetic drugs — PEPT1 is the principal apical peptide transporter in intestinal epithelium. Overexpression is useful for testing peptide-substrate transport or for studying PEPT1-targeted prodrugs (valacyclovir, valganciclovir). For oral drug bioavailability research, the EDITGENE PEPT1 Knockout in Huh-7 enables study of peptide transporter pharmacology — though Huh-7 has hepatic rather than intestinal origin. PEPT2 (SLC15A2) paralog expression analysis aids interpretation. Rescue with wild-type or transport-deficient PEPT1 is the standard specificity control for PEPT1-targeted prodrug research. The knockout is valuable for PEPT1-targeted drug delivery strategy development.

Primary applications: • Peptide uptake: ³H-dipeptide (Gly-Sar, others) uptake assays to quantify PEPT1 transport activity. • Peptide-mimetic prodrug uptake: valacyclovir, valganciclovir, and other PEPT1-substrate prodrug uptake studies. • pH-dependence: PEPT1 transport is proton-coupled — assays under varied extracellular pH characterize transport mechanism. • Substrate scope: assessment of clinically relevant PEPT1 substrates (β-lactam antibiotics, ACE inhibitors). EDITGENE recommends this model for researchers investigating intestinal peptide transport, oral drug bioavailability, and PEPT1-targeted prodrug development.

Yes. PEPT1 rescue experiments are well-established for peptide transport research: • Construct design: use a codon-modified SLC15A1 sequence with a small C-terminal tag (FLAG, HA). PEPT1 has 12 transmembrane domains with apical sorting determinants. • Surface localization validation: confirm plasma membrane localization before transport assays. • Transport-deficient rescue: substrate-binding pocket mutations enable structure-function studies. • Functional readout: rescue should restore dipeptide uptake activity and pH-dependent peptide-mimetic prodrug uptake. Huh-7 transduces efficiently with lentivirus and supports stable rescue line generation.